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Figure 3


Fig. 3. Ca2+ is required for STAg-induced production of IL-12. Macrophages pre-treated with normal buffer (black bars) or BAPTA-AM (15 µM) in Ca2+ free/EGTA (white bars) were stimulated with STAg (50 µg/ml) for 5 hours in normal media or Ca2+ free EGTA medium, respectively. mRNA was isolated and reverse transcribed to cDNA, which was used in real-time PCR reactions for IL-12p35 and IL-12p40. All amplifications were conducted in triplicate and results are normalized to endogenous 18S and expressed as fold change in mRNA relative to untreated controls for each condition, according to the {Delta}{Delta}Ct method. Experiments were repeated at least three times with similar results. The data presented are from a representative experiment. *P≤0.01 by Student's t-test.





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